Xenobiotic oxidation by cytochrome P-450-enriched extracts of Streptomyces griseus

Crude extracts of Streptomyces griseus grown on soybean flour-enriched medium contain high levels of cytochrome P-450. The cytochrome P-450-enriched fractions, obtained by ammonium sulfate fractionation (30-50% saturation), catalyze the NADPH-dependent oxidation of a variety of xenobiotics when complemented with both spinach ferredoxin:NADP+ oxidoreductase and spinach ferredoxin. Reactions observed are aromatic, benzylic and alicyclic hydroxylations, O-dealkylation, non-aromatic double bond epoxidation, N-oxidation and N-acetylation.     

25-Hydroxylation of 1 alpha-hydroxyvitamin D-3 in rat and human liver

1 alpha-Hydroxyvitamin D-3 25-hydroxylase activity was measured in subcellular fractions of rat and human liver. The formation of 1,25-dihydroxyvitamin D-3 was determined by high pressure liquid chromatography. In rat liver 1 alpha-hydroxyvitamin D-3 25-hydroxylase activities were found in the purified nuclei, the heavy mitochondrial fraction and the microsomal fraction. The enrichment of 25-hydroxylase activity was highest in the heavy mitochondrial fraction. With this fraction a minimum amount (about 0.5 mg) of protein was required before formation of 1,25-dihydroxyvitamin D-3 could be detected. Above this amount the reaction was linear with amount of protein up to at least 2 mg/ml. The reaction was also linear with time up to 60 min. An apparent Km value of 2 X 10(-5) M was found. The mitochondrial 25-hydroxylase was stimulated by addition of cytosolic protein or bovine serum albumin. The degree of stimulation was dependent on the amount of mitochondrial protein present in the incubation mixture. Maximal stimulation was seen with 0.2 mg/ml of either protein in the presence of 0.5 mg mitochondrial protein. The stimulating effect remained after heating the protein for 5 min at 100 degrees C. The cytosolic protein did not stimulate a reconstituted mitochondrial 1 alpha-hydroxyvitamin D-3 25-hydroxylase. The mitochondrial vitamin D-3 25-hydroxylase was inhibited both by cytosolic protein and by bovine serum albumin. Human liver revealed only one 1 alpha-hydroxyvitamin D-3 25-hydroxylase activity located to the heavy mitochondrial fraction. The results are in agreement with previous studies on the localization of vitamin D-3 25-hydroxylase in rat and human liver. The difference in localization of the 25-hydroxylase between rat and human liver implies that studies on the regulation of the microsomal 25-hydroxylase in rat liver may not be relevant to the situation in human liver.     

A new longirostrine dyrosaurid (Crocodylomorpha,Mesoeucrocodylia) from the Paleocene of north‐eastern Colombia: biogeographic and behavioural implications for New‐World Dyrosauridae

Abstract: Fossils of dyrosaurid crocodyliforms are limited in South America, with only three previously diagnosed taxa including the short‐snouted Cerrejonisuchus improcerus from the Paleocene Cerrejón Formation of north‐eastern Colombia. Here we describe a second dyrosaurid from the Cerrejón Formation, Acherontisuchus guajiraensis gen. et sp. nov., based on three partial mandibles, maxillary fragments, teeth, and referred postcrania. The mandible has a reduced seventh alveolus and laterally depressed retroarticular process, both diagnostic characteristics of Dyrosauridae. Acherontisuchus guajiraensis is distinct among known dyrosaurids in having a unique combination of craniomandibular characteristics, and postcranial morphology that suggests it may have occupied a more placid, fluvial habitat than most known Old‐World dyrosaurids. Results from a cladistic analysis of Dyrosauridae, using 82 primarily cranial and mandibular characters, support an unresolved relationship between A. guajiraensis and a combination of New‐ and Old‐World dyrosaurids including Hyposaurus rogersii, Congosaurus bequaerti, Atlantosuchus coupatezi, Guarinisuchus munizi, Rhabdognathus keiniensis and Rhabdognathus aslerensis. Our results are consistent with an African origin for Dyrosauridae with multiple dispersals into the New World during the Late Cretaceous and a transition from marine habitats in ancestral taxa to more fluvial habitats in more derived taxa.     

Biochemical analysis and synergistic suppression of indoxacarb resistance in Plutella xylostella L.

Indoxacarb was treated to Plutella xylostella for 10 generations to develop a resistant strain and biochemical analysis of indoxacarb resistance in different tissues of P. xylostella was carried out. Biochemical analysis found maximum esterase activity in gut homogenates of indoxacarb resistant strains followed by whole body and cuticle homogenates. In gut homogenates of indoxacarb resistant strains, maximum increase in esterases was found as compared to the unselected strain. Acetylcholineesterase activity was higher in head homogenates of the resistant strain than in the unselected strain. Glutathione-S-transferase activity was highest in whole body homogenates. However, maximum increase was found in gut homogenates of indoxacarb resistant strains over the unselected. Induced resistance was suppressed using known synergists. Maximum synergism occurred using diethyl-maleate (DEM), followed by triphenyl phosphate (TPP).